CLASSIFICATION TESTS FOR HYDROXYL- AND CARBONYL- CONTAINING COMPOUNDS

Kathleen Anne Francisco, Ma. Casey Louisse Garcia, Aimee Dianne Hermoso, Frances Geraldine Ibale, Rigel Larga Group IV, 2EPharmacy, Faculty of Pharmacy, University of Santo Tomas

ABSTRACT

Hydroxyl and Carbonyl containing compounds belong to different functional groups in Organic Chemistry thus, they each have distinct physical and chemical properties. In this experiment, series of tests were conducted to differentiate the two functional groups. Included in these tests were solubility of alcohols in water, Lucas test, Chromic Acid test also known as Jones Oxidation, 2,4-Dinitrophenylhydrazone or 2,4DNP test, Fehlings test, Tollens Silver Mirror test and Iodoform test. Sample compounds used for this experiment were ethanol, n-butyl alcohol, sec-butyl alcohol, tert-butyl alcohol, benzyl alcohol, n-butyraldehyde, benzaldehyde, acetone,

acetophenone, isopropyl alcohol, and acetaldehyde. Based on the results obtained, ethanol, sec-butyl alcohol, and tert-butyl alcohol are miscible in 1, 28, and 5 drops of water respectively. While n-butyl alcohol is slightly miscible with 40 drops of water and benzyl alcohol is immiscible with water. For Lucas test, sec-butyl alcohol and tert-butyl alcohol formed a cloudy suspension or two distinct layers. Green coloration of the solution was observed in n-butyl alcohol, sec-butyl alcohol, n-butyraldehyde, and benzaldehyde in the Chromic Acid test. For 2,4-DNP test, acetone, acetaldehyde, nbutyraldehyde, benzaldehyde, and acetophenone formed a yellow precipitate. A brick-red precipitate was observed in acetaldehyde, n-butyraldehyde, and benzaldehyde in the Fehlings test. Silver mirror formation was present in n-butyraldehyde, acetaldehyde and benzaldehyde for the Tollens Silver Mirror test. For the Iodoform test, there was the formation of yellow precipitate on acetaldehyde, acetone, isopropyl alcohol, and acetophenone.

INTRODUCTION
According to Helmenstine, in both laboratory and biochemical transformations of organic compounds. They also play a central role in the interconversion of organic functional groups (Brown & Poon, 2014). Alcohols are divided into primary (1), secondary (2) and tertiary (3) compounds. This classification is based on the type of carbon to which the hydroxyl group is attached (Kelly). A primary alcohol is attached to a carbon containing only one alkyl group, a secondary alcohol is attached to a carbon containing two alkyl grouos and a tertiary alcohol is attached to a carbon containing three alkyl groups. Some of the sample compounds belonging to this group are: n-butyl alcohol (1 alcohol), ethanol (1 alcohol), sec-butyl alcohol (2 alcohol), tert-butyl alcohol (3 alcohol), isopropyl alcohol (2 alcohol) and benzyl alcohol (1 alcohol). Organic compounds having a C=O functional group are named

Organic Chemistry is a discipline that is concerned with the study of compounds containing Carbon that is chemically bonded to Hydrogen. Organic compounds are abundant around us in our foods, flavors, and fragrances; in our medicines, toiletries, and cosmetics; in our plastics, films, fibers, and resins; in our paints and varnishes; in our glues and adhesives; and of course, in our bodies and in all living things (Brown & Poon, 2014). The properties and

reactivities of organic molecules are largely determined by the kinds of functional groups a given molecule possesses. They contain either

heteroatoms like halogens, nitrogen or oxygen, or carbon-carbon double or triple bonds (Suggs, 2002). Compounds with the same functional group undergo similar chemical reactions (Hornback, 2006). In addition to these, functional groups are also the units by which we can divide organic compounds into families and are the basis for naming each family (Brown & Poon, 2014). Alcohol is an organic compound that has the functional group OH (hydroxyl) group that is attached to an alkyl group thus, its general formula is R-OH. They are particularly important

grouped together as carbonyl containing compounds. This includes carboxylic acid and derivatives, ketones and

aldehydes. Aldehydes have at least one hydrogen attached to the carbonyl carbon thus, its general formula is RCOH. Ketones on the other hand have no hydrogen attached to the carbonyl

carbon, only carbon containing R group thus, its general formula is RCOR (Bettelheim & Landesberg, 2010).

sodium hydroxide), Tollens reagent (5% silver nitrate, and 2% 5% sodium

hydroxide,

ammonium

Aldehydes and Ketones are found widely distributed in the animal and plant kingdoms. They are responsible for the sweet and penetrating aroma of fruit and flowers. They are also

hydroxide), 5% sodium hypochlorite, iodoform test reagent(10% potassium iodide and 5% sodium hypochlorite), and 2,4-dinitrophenylhydrazine. The

sample compounds used were ethanol, n-butyl alcohol, sec-butyl alcohol, tertbutyl alcohol, benzyl alcohol, n-

components of larger biomolecules such as cellulose and nucleic acids (Palleros, 2000). Sample compounds belonging to this functional group are: n-

butyraldehyde, benzaldehyde, acetone, acetophenone, isopropyl alcohol, and acetaldehyde. SOLUBILITY OF ALCOHOLS IN

butyraldehyde, benzaldehyde, acetone, acetophenone and acetaldehyde. The objectives of this

WATER Ethanol, n-butyl alcohol, secbutyl alcohol, tert-butyl alcohol, and benzyl alcohol were used in this solubility test. Ten drops of each were put in different test tubes and 20 drops of water was added initially. Shaking the mixture after addition of water is

experiment are to : (1) distinguish whether a compound is hydroxyl or carbonyl containing , (2) differentiate the three types of alcohols, (3)

differentiate aldehydes from ketones and (4) explain the mechanisms involved in the differentiating tests.

MATERIALS AND METHODS

Materials In this experiment, the

important to ensure thorough mixing of the two liquids. Adding successive volumes of water is done until a homogenous dispersion is achieved. The total volume of water added was noted down. LUCAS TEST The researchers first prepared the Lucas reagent. This was done by dissolving 16grams of anhydrous zinc

researchers made used of different chemical reagents. First is the Lucas reagent (anhydrous zinc chloride and concentrated sulfuric acid), chromic acid reagent, 95% ethanol, Fehlings A (hydrated copper (II) sulfate), Fehlings B (Potassium sodium tartrate and

chloride in ten milliliters of concentrated sulfuric acid. The mixture was then allowed to cool. After this, two to three drops of each of the sample (n-butyl alcohol, sec-butyl alcohol, and tert-butyl alcohol) were added to one milliliters of the freshly prepared lucas reagent. The mixture was shaken vigorously and allowed to stand at room temperature. CHROMIC ACID TEST (JONES OXIDATION) One drop of each of the sample compounds (n-butyl alcohol, sec-butyl alcohol, tert-butyl alcohol, n-

sample was added to five drops of 95% ethyl alcohol. Three drops of 2.4dinitrophenylhydrazine was then added. FEHLINGS TEST The reagent was first prepared by mixing Fehlings A and Fehlings B. Fehlings A was obtained by dissolcing seven grams of hydrated copper (II) sulfate in 100mililiters of water. Fehlings B on the other hand was prepared by dissolving 35 grams of potassium sodium tartrate and ten grams of sodium hydroxide in 100 mililiters of water. After this, three drops of each of the sample (acetaldehyde, acetone, n-

butyraldehyde, benzaldehyde, acetone and acetophenone) was added to one milliliter of acetone. Two drops of 10%aqueous potassium permanganate and five drops of 6M sulfuric acid were then added to the solution. 2,4-dinitrophenylhydrazone test This test was performed on acetone, acetaldehyde, n-butyraldehyde, benzaldehyde and acetophenone. The reagent was first prepared by adding a solution of three grams of 2,4-

butyraldehyde,

benzalehyde

and acetophenone) was added to one milliliter of the freshly prepared Fehlings reagent. The samples were then placed in a beaker of boiling water and changes were observed after 1015minutes. TOLLENS TEST The Tollens reagent was first prepared. This was done by mixing two milliliters of 5% silver nitrate solution and two drops of 5% sodium hydroxide. The mixture was mixed thoroughly and was added drop by drop of 2% ammonium hydroxide until all the precipitate was dissolved. After the SILVER MIRROR

dinitrophenylhydrazine to 15 milliters of concentrated sulfuric acid. While

stirring, 20mililiters of water and 70 mililiters of 95% ethanol were then added. The solution was mixed and filtered. A drop of each of the liquid

preparation of the reagent, two drops of each of the samples (acetaldehyde, nbutyraldehyde, benzaldehyde, acetone, and acetophenone) was mixed with 20 drops of the freshly prepared reagent. The mixture was then allowed to stand for ten minutes. The test sample without a reaction was subjected in a beaker of warm water for five minutes. Changes were noted down. IODOFORM TEST Two drops of each of

dissolve non-polar solute. The polarity of organic molecules is determined by polar bonds due to the presence of electronegative atoms like Nitrogen, Fluorine and Oxygen in polar functional groups such as amines and alcohols (http://www.chem.ucalgary.ca/courses/3 51/laboratory/351expt_01_solubility.pdf , Retrieved September 11, 2013). Alcohol has a functional group of OH. Thus, they are polar molecules and can form dipole-dipole called

intermolecular

attraction

acetaldehyde, acetone, acetophenone, benzaldehyde, and isopropyl alcohol were placed in different test tubes. The test samples were then added with 20 drops of 10% potassium iodide solution and 20 drops of 5% sodium

hydrogen bonding (Brown & Poon, 2014).

hypochlorite. Observations were then noted down.

Figure 1. General formula of Alcohol Water is known as the universal solvent. This has to do with the polarity

RESULTS AND DISCUSSION

According to Merriam-Webster dictionary, solubility is the amount of a substance that will dissolve in a given amount of another substance. At the molecular level, solubility is controlled by intermolecular forces. A general rule of like dissolves like is applied in solubility. This rule is based on polarity of the systems. Polar solvents dissolve polar solutes while non-polar solvents

of each water molecule. The hydrogen side of each water molecule carries a slight positive electric charge, while the oxygen side carries a slight negative electric charge (Helmenstine). This

accounts for the ability of water to dissolve substances.

Figure 2. Structure of Water

There are many factors that can affect the solubility of organic

Ethanol or Ethyl Alcohol is a clear, colorless primary alcohol.

compounds in water. As the percentage of the hydrocarbon increases, solubility in water decreases. Increase in Figure 3. Structure of Ethanol It only takes one drop of water to produce a homogenous suspension of the mixture. Aside from its polar group, the short chain of carbon atoms

molecular weight decreases alcohol solubility in water. Another one,

compounds having a branched chain is more soluble than the corresponding straight-chain compound. Position of the functional solubility (http://opencourseware.kfupm.edu.sa/col leges/cs/chem/chem303/files%5C3Lecture_Notes_CHEM303_(Chapter_5).pdf, September 11, 2013). For this test, Ethanol, n-butyl alcohol, sec-butyl alcohol, tert-butyl alcohol, and benzyl alcohol were used. Table 1. Solubility of alcohol in Water Alcohol
Amount of water needed to produce a homogenous suspension

group

also

affects

its

contributes to its high solubility. Thus, it is miscible in water.

Retrieved Figure 4. Structure of n-butyl alcohol n-butyl alcohol or n-butanol is also a primary alcohol. It takes 40 drops of water to produce a homogenous suspension of the mixture. Thus, it is only slightly miscible in water. The long chain of carbon atoms contributes to its decreased solubility in water. Miscible Slightly miscible Miscible Miscible Figure 5. Structure of sec-butyl alcohol Immiscible According to its structure, secbutyl alcohol or 2-butanol is a

Solubility

Ethanol n-butyl alcohol sec-butyl alcohol tert-butyl alcohol benzyl alcohol

1 drop 40 drops 28 drops 5 drops ________

secondary alcohol. It has the same number of carbon atoms with n-butyl alcohol but it only takes 28 drops of water to produce a homogenous

Benzyl alcohol is a primary alcohol. Based on the experiment, it does not form a homogenous suspension with water. It is because of the presence of the benzene ring. Benzene ring possesses a high degree of unsaturation because of the presence of double bonds and double bonds are harder to break than single bonds (Zumdahl, 2012). Thus, it has a lower possibility of reacting with water. It is therefore immiscible in water. Table 2. Results of the Lucas Test Test Samples Lucas Test No visible positive result Formation of a cloudy suspension within one minute Formation of a cloudy suspension within five seconds

suspension of the mixture. It is because of the position of its functional group. The functional group is attached to its second carbon atom thus, it facilitates faster and easier breaking of the bond of the structure. Therefore, it is miscible in water.

Figure 6. Structure of tert-butyl alcohol tert-butyl alcohol is a tertiary alcohol. It has the same number of carbon atoms with that of n-butyl alcohol and sec-butyl alcohol but it only takes five drops of water to produce a homogenous suspension of the mixture. It is because the more branched the structure, the higher its solubility in water. Thus, it is miscible.

n-butyl alcohol sec-butyl alcohol

tert-butyl alcohol

Figure 7. Structure of Benzyl alcohol

Figure 8. Reaction Mechanism of Lucas Test Lucas test is used to distinguish between primary, secondary and tertiary

alcohols.

According the

to reaction

the that

sec-butyl alcohol

Green

coloration

Organicchem.org,

of solution tert-butyl alcohol No visible positive result n-butyraldehyde Green coloration

occurs in the Lucas test is an SN1 nucleophilic substitution. When the lucas reagent is added to the alcohol, the proton from hydrochloric acid will protonate the hydroxyl group of alcohol forming a good leaving group of water. This will result in the formation of a carbocation and the release of chloride from hydrochloric acid. The water will then be released and the chloride ion will be attracted to the carbocation. This will result in the formation of an alkyl halide. The positive visible of which is the formation of a cloudy suspension or two layers which is the insoluble alkyl chloride. Tertiary alcohols like tert-butyl alcohol immediately reacts with the lucas reagent. This is because the speed of this reaction is directly proportional to the amount of energy required to form the carbocation (chemuscb.edu). Thus, secondary alcohols like the sec-butyl alcohol react slower than tertiary ones. The primary alcohol like the n-butyl alcohol has the least possibility of reacting with lucas reagent.

of solution benzaldehyde Green coloration

of solution acetone No visible positive result acetophenone No visible positive result

Figure 9. The Chromic Acid Reaction The reagent of this test is chromium trioxide in sulfuric acid. Primary alcohols and aldehydes are oxidized to carboxylic acids using this test (Dr. Pahlavan/Cherif). Secondary alcohols can also be oxidized to ketones. They will react to the yellow-orange Jones reagent that contains chromium

Table 3. Results of the Chromic Acid Test Test Samples n-butyl alcohol Chromic Acid Test Green coloration of solution

IV ion by nucleophilic addition which will be reduced to chromium III which is green (Dr. Pahlavan/Cherif). Thus, nbutyl alcohol, sec-butyl alcohol, n-

butyraldehyde,

and

benzaldehyde

These are yellow-red precipitates that can be used to characterize aldehydes and ketones. Other functional groups such as esters and amides do not react with this reagent (Palleros, 2000). This reaction can be described with as a two

showed a positive visible result on this test because they are classified as primary alcohol, secondary alcohol and aldehydes respectively.

Table 4. Results of 2,4dinitrophenylhydrazone test Test Samples Acetone 2,4-DNP test Formation of

condensation

reaction,

molecules joining together with loss of water. It elimination is also called : addition-

reaction

nucleophilic

addition of amino group to the carbonyl group of either aldehyde or ketone, followed by the removal of water (Dr. Pahlavan/Cherif). acetaldehyde, Thus, acetone,

yellow precipitate Acetaldehyde Formation of

yellow precipitate Benzaldehyde Formation of

n-butyraldehyde,

yellow precipitate Acetophenone Formation of

benzaldehyde, and acetophenone formed a yellow precipitate upon the addition of the reagent. Table 5. Results of Fehlings Test Test Samples Acetaldehyde Fehlings test Formation of brick-red precipitate Formation of brick-red precipitate No visible positive result Benzaldehyde Formation of brick-red precipitate No visible positive result

yellow precipitate n-butyraldehyde Formation of

yellow precipitate

n-butyraldehyde

Acetone Figure 10. Formation of 2,4dinitrophenylhydrazone Aldehydes and ketones react with 2,4-dinitrophenylhydrazine, under acidic conditions, to give 2,4Acetophenone

dinitrophenylhydrazones. The reaction proceeds by nucleophilic addition.

Figure 11. The Fehlings Test Reaction In this reaction, copper II in an alkaline solution is the oxidizing agent. It is also used to prevent the

Figure 12. The Tollens Test Reaction Aldehydes react with a solution of silver ion in ammonia to give carboxylic acids and metallic silver. Ketones on the other hand are not oxidized by this reagent (Palleros, 2000). Thus, acetone and acetophenone did not show a visible positive result on this test. In this test, the aldehyde reduces silver to metallic In silver

precipitation of copper hydroxide by complexing it with tartrate. Aldehydes can reduce the Fehlings reagent. Thus, acetaldehyde, n-butyraldehyde, and

benzaldehyde showed a positive visible result. This is shown by the formation of green solution and finally reddish precipitate of the copper oxide. Other functional groups such as hydroxyl ketones and carbohydrates are also capable of reacting with the Fehlings reagent (Palleros, 2000).

(myweb.brooklyn).

addition,

Tollens reagent should be prepared immediately before use and should never be stored because it forms explosive silver nitrate upon standing (Palleros, 2000).

Table 6. Results of Tollens Silver Mirror Test Test Samples Acetaldehyde Benzaldehyde Acetone n-butyraldehyde Acetophenone Tollens Test Formation of silver mirror Formation of silver mirror No visible positive result Formation of silver mirror No visible positive result

Table 7. Results of the Iodoform test Test Samples Acetaldehyde Iodoform test Formation of

yellow precipitate Acetone Formation of

yellow precipitate Acetophenone Formation of

yellow precipitate Benzaldehyde No formation of visible result Isopropyl alcohol Formation of positive

yellow precipitate

attacks the carbonyl, which has made more electron deficient by the -

CI3substituent. With the three electron withdrawing iodine atoms attached, the CI3 is a good leaving group

(myweb.brooklyn). Figure 13. The Iodoform Reaction

This test identifies the presence of a methyl ketone functional group. They are oxidized to carboxylic acids. Acetaldehydes also give a positive visible result in this test owing to its structural similarity to methyl ketones. Secondary alcohols can also be oxidized to methyl ketones to give a positive result in this test (Dr. Pahlavan/Cherif). Thus, acetaldehyde, acetone,

REFERENCES :
BOOKS Bettelheim, F. & Landesberg, J. (2010). Laboratory Experiments for Introduction Organic, and to General,

Biochemistry.

Cengage Learning Brown, W. & Poon, to T. (2014). Organic

Introduction

acetophenone and isopropyl alcohol gives a positive visible result which is the formation of a yellow precipitate. In the basic reaction conditions, the proton that is next to the carbonyl (the proton) is removed. This makes the carbon into a nucleophile that attacks a molecule of iodine in the second step. The presence of one iodine atom on the -carbon increases the acidity of the remaining hydrogens. Each one is also removed by the base to make a nucleophile that attacks more iodine molecules until all three hydrogens are replaced by iodine. The hydroxide ion

Chemistry. Bowling Green State University : John Wiley & Sons Hornback, J. (2006). Organic Learning,

Chemistry.Thomson Inc. Palleros, D. (2000).

Experimental

Organic Chemistry. University of California : John Wiley & Sons Snuggs, J. (2002). Organic Chemistry. New York : Barrons

Educational Series, Inc.

Zumdahl, S. (2012). Chemistry An Atoms First Approach.

United States of America : Brooks/Cole. ONLINE http://myweb.brooklyn.liu.edu/swatsn/ Site/Laboratory_Manuals_fils/Exp13.pd f, Retrieved September 11, 2013

http://opencourseware.kfupm.edu.sa/coll eges/cs/chem/chem303/files%5C3Lecture_Notes_CHEM303_(Chapter_5).pdf, September 11, 2013 http://chemistry.about.com/od/chemis tryglossary/a/organicchemdef.htm, Retrieved September 10, 2013 Pahlavan & Cherif (2013). Identification of Aldehydes and Ketones. Organic Chemistry II. Retrieved from http://swc2.hccs.edu/pah avan/2425L6.pdf Retrieved on September 10, 2013 Retrieved